Time- and concentration-dependent cytotoxicity of antibiotics used in endodontic therapy

OBJECTIVE: New drugs have to be assessed in endodontic therapy due to the presence of microorganisms resistant to therapeutic procedures. Thus, this study evaluated the time- and concentration-dependent cytotoxicity of different antibiotics used in endodontic therapy. MATERIAL AND METHODS: Human gingival fibroblasts were treated and divided into the following experimental groups: Group I - control; Group II - ciprofoxacin hydrochloride; Group III - clyndamicin hydrochloride; and Group IV - metronidazole. Each drug was used at concentrations of 5, 50, 150, and 300 mg/L for 24, 48, 72, and 96 h. Cytotoxicity was evaluated by the MTT assay [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] and spectrophotometric reading of ELISA plates. The results were analyzed by BioEstat 4.0 software using Kruskal-Wallis and Dunn's tests at a signifcance level of 5%. Cell viability was assessed for the different concentrations and times. RESULTS: All drugs presented dose-dependent cytotoxicity. Concentrations of 5 and 50 mgjL produced viable fibroblasts at all experimental times in all groups. CONCLUSIONS: Cell viability at 24 h was greater than in the other experimental times. Comparison between the same concentrations of antibiotics at different times showed that metronidazole presented the highest cell viability at 72 and 96 h compared to the other antibiotics, whereas clyndamicin hydrochloride showed higher cell viability at 72 h than ciprofoxacin hydrochloride.

Microbial culture and checkerboard DNA-DNA hybridization assessment of bacteria in root canals of primary teeth pre- and post-endodontic therapy with a calcium hydroxide/chlorhexidine paste

Aim. To investigate the root canal microbiota of primary teeth with apical periodontitis and the in vivo antimicrobial effects of a calcium hydroxide/chlorhexidine paste used as root canal dressing. Design. Baseline samples were collected from 30 root canals of primary teeth with apical periodontitis. Then, the root canals were filled with a calcium hydroxide paste containing 1% chlorhexidine for 14 days and the second bacteriologic samples were taken prior to root canal filling. Samples were submitted to microbiologic culture procedure to detect root canal bacteria and processed for checkerboard DNA-DNA hybridization. Results. Baseline microbial culture revealed high prevalence and cfu number of anaerobic, black-pigmented bacteroides, Streptococcus, and aerobic microorganisms. Following root canal dressing, the overall number of cfu was dramatically diminished compared to initial contamination (P < 0.05), although prevalence did not change (P > 0.05). Of 35 probes used for checkerboard DNA-DNA hybridization, 31 (88.57%) were present at baseline, and following root canal dressing, the number of positive probes reduced to 13 (37.14%). Similarly, the number of bacterial cells diminished folowing application of calcium hydroxide/chlorhexidine root canal dressing (P = 0.006). Conclusion. Apical periodontitis is caused by a polymicrobial infection, and a calcium hydroxide/chlorhexidine paste is effective in reducing the number of bacteria inside root canals when applied as a root canal dressing.

Evaluation of periapical changes following endodontic therapy: digital subtraction technique compared with computerized morphometric analysis

Objectives: The aim of this study was to compare two methods for the evaluation of periapical lesion changes following endodontic therapy (digital subtraction technique and morphometric analysis) by outlining the radiolucent area.Methods: 13 human anterior teeth with pulp necrosis and chronic periapical lesions were used. Periapical radiographs were taken immediately after endodontic therapy (0) and then 2 months, 4 months and 6 months post treatment, using an intraoral radiographic film holder stabilized with impression material. The films were processed in a standard manner and the digitized images were submitted to digital subtraction using Adobe Photoshop 6.0. New bone formation or bone resorption areas were then measured. In the morphometric analysis, the periapical lesions were outlined using VixWin 2000 and the area (in square millimetres) was recorded. The obtained data were submitted to agreement analysis for comparison of the two techniques.Results: There was no correlation between the areas of radiographic changes detected by digital subtraction and periapical lesion outline (r=0.02-0.45). The new bone formation areas observed by digital subtraction presented higher values, with bone changes being especially evident in the 2 month follow-up radiographs, which suggests a higher sensitivity for this method.Conclusions: Both methods are suitable for the evaluation of periapical lesion changes, but the digital subtraction technique is more sensitive for detecting radiographic periapical changes. Dentomaxillofacial Radiology (2009) 38, 438-444. doi: 10.1259/dmfr/53304677

A comparison of one versus two appointment endodontic therapy in dogs' teeth with apical periodontitis

This study was conducted to observe the healing process in dogs' teeth with apical periodontitis after root canal treatment in one or two appointments. Premolars and anterior dogs' teeth had their root canals opened to the oral environment for 6 months before being treated. After root canal negotiation they were filled by the lateral condensation technique with gutta-percha points and Sealapex in one appointment or after a dressing with calcium hydroxide for 7 and 15 days. Six months after the treatment the animals were killed and the tissues prepared for histomorphological analysis. Scores attributed to the different histomorphological events were submitted to statistical analysis, which resulted in ranking the experimental groups from the best to the worst in the following way: (a) calcium hydroxide 14 days; (b) calcium hydroxide 7 days; and (c) one appointment. It was concluded that the use of a calcium hydroxide dressing helps to achieve better results (p < 0.01) than the treatment in one appointment.

Proinflammatory Activity Of Primarily Infected Endodontic Content Against Macrophages After Different Phases Of The Root Canal Therapy.

This study investigated the presence of target bacterial species and the levels of endotoxins in teeth with apical periodontitis. Levels of inflammatory mediators (interleukin [IL]-1β and tumor necrosis factor [TNF]-α) were determined after macrophage stimulation with endodontic content after different phases of endodontic therapy using different irrigants. Thirty primarily infected root canals were randomly assigned into 3 groups according to the irrigant used for root canal preparation (n = 10 per group): GI: 2.5% sodium hypochlorite, GII: 2% chlorhexidine gel, and GIII (control group): saline solution. Root canal samples were taken by using paper points before (s1) and after root canal instrumentation (s2), subsequently to 17% EDTA (s3), after 30 days of intracanal medication (Ca[OH]2 + saline solution) (s4), and before root canal obturation (s5). Polymerase chain reaction (16S recombinant DNA) and limulus amebocyte lysate assay were used for bacterial and endotoxin detection, respectively. Macrophages were stimulated with the root canal contents for IL-1β/TNF-α measurement using enzyme-linked immunosorbent assay. Porphyromonas gingivalis (17/30), Porphyromonas endodontalis (15/30), and Prevotella nigrescens (11/30) were the most prevalent bacterial species. At s1, endotoxins were detected in 100% of the root canals (median = 32.43 EU/mL). In parallel, substantial amounts of IL-1β and TNF-α were produced by endodontic content-stimulated macrophages. At s2, a significant reduction in endotoxin levels was observed in all groups, with GI presenting the greatest reduction (P < .05). After a root canal rinse with EDTA (s3), intracanal medication (s4), and before root canal obturation (s5), endotoxin levels reduced without differences between groups (P < .05). IL-1β and TNF-α release decreased proportionally to the levels of residual endotoxin (P < .05). Regardless of the use of sodium hypochlorite or CHX, the greatest endotoxin reduction occurs after chemomechanical preparation. Increasing steps of root canal therapy associated with intracanal medication enhances endotoxin reduction, leading to a progressively lower activation of proinflammatory cells such as macrophages.

Novel biomaterials for regeneration of periapical bone defects in endodontic and implant therapy

Endodontic-related periapical bone defects are a common occurrence in the global populations. Considering the number of root canal treatments performed annually, new strategies and new biomaterials for the management of these bone defects will be important and highlight the need for continued research and development in endodontic field. The present PhD thesis have several objectives and is divided into two main sections: one focused on in vitro and laboratory research and the other on clinical in vivo investigations. The first part, focused on laboratory and in vitro research, investigated 2 main topics: • the microbial communities of apical periodontitis to evaluate the predominant bacterial using 16sr DNA-targeted Nanopore sequencing; • the physical-chemical properties of innovative premixed calcium-silicate based bioceramic sealers for endodontic therapy; The second part, focused on in vivo clinical studies, investigated 2 main topics: • the clinical application of premixed calcium-silicate-based sealers. Ethical committee approval was obtained in 2 separate in vivo studies. The first one is a prospective cohort study with a two-year follow-up where the test group was compared with a control group (considered the gold standard). The second is a pilot prospective cohort study with a 12-month follow-up which set the foundation for a subsequent randomized investigation. Thanks to these investigations, we validated a new technique that innovatively associates a warm obturation technique with calcium-silicate-based sealers. Historically, these sealers were only used with cold techniques. This investigation highlights the possibility for wider utilization and improvements in endodontic techniques. • The outcome of 2 different types of implants characterized by different surface treatments and placed with different techniques. The marginal bone level and periodontal parameters were evaluated with a follow-up of 4 and 10 years. This Ph.D thesis is based on a compilation of published papers I have done during my three-year PhD program.

Evaluation Of Photodynamic Therapy Using A Diode Laser And Different Photosensitizers Against Enterococcus Faecalis.

Photodynamic therapy (PDT) has been proven to be effective in disinfecting root canals. The aim of this present study was to evaluate the effects of PDT on the viability of Enterococcus faecalis using methylene blue (MB) and malachite green (MG) as photosensitizers. Solutions containing E. faecalis (ATCC 29212) were prepared and harvested by centrifugation to obtain cell suspensions, which were mixed with MB and MG. Samples were individually irradiated by the diode laser at a distance of 1mm for 30, 60, or 120 seconds. Colonyforming units (CFU) were determined for each treatment. PDT for 60 and 120 seconds with MG reduced E. faecalis viability significantly. Similar results were obtained when MB was used as photosensitizer. PDT using MB and MG have antibacterial effect against E. faecalis, showing potential to be used as an adjunctive antimicrobial procedure in endodontic therapy.

Diagnosis and treatment of odontogenic cutaneous sinus tracts of endodontic origin: three case studies

To describe three cases of extraoral sinus tracts, related to infected teeth, which were initially misdiagnosed as skin lesions and inappropriately treated. The extraoral sinus tracts were initially misdiagnosed as skin lesions. Dermatological surgery was performed and antibiotics prescribed but the lesions did not resolve. Then, a dental cause was sought, and identified. Endodontic intervention resulted in resolution of the problem, confirming the initial misdiagnosis. center dot Dermatologists and other medical practitioners should be aware that dental extraoral sinus tracts can be confused with skin lesions. center dot A dental aetiology, as part of a differential diagnosis, should be kept in mind with oro-facial skin lesions. center dot If an extraoral sinus tract is of endodontic origin, then elimination of infection through effective endodontic treatment will lead to resolution of the sinus tract. center dot Early correct diagnosis can prevent unnecessary and ineffective antibiotic therapy and/or surgical intervention.

Use of computerized tomography for diagnosis and follow-up after endodontic surgery: clinical case report with 8 years of follow-up

Computerized tomography (CT) is a valuable tool for diagnosis and planning in conventional and surgical endodontic therapy. This case report describes the use of CT in the diagnosis of a periapical lesion undetected by periapical radiography in the mandibular molar area. The CT also showed a possible mesial root perforation associated with the lesion. Following CT, surgical planning, periradicular curettage, and sealing of the root perforation were performed. Eight years after surgery, cone beam CT revealed periapical bone repair. Computerized tomography can be an important resource for diagnosis and planning in conventional and surgical endodontic therapy, as well as for evaluation of post-treatment bone repair. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2010; 109: 629-633)

Use of cone-beam tomography and digital subtraction radiography for diagnosis and evaluation of traumatized teeth treated with endodontic surgery and MTA. A case report

The aim of this paper was to report the diagnosis and evaluation of periradicular bone repair by using computed tomography and digital subtraction radiography in an endodontic surgery case treated with mineral trioxide aggregate (MTA). The patient had local swelling and fistula 7years after trauma in the upper jaw. Periapical lesion on tooth #12 and root resorption of tooth #11 were detected radiographically. Endodontic therapy and placement of intracanal medication were carried out. After 3months, with no improvement in the clinical signs and symptoms, computed tomography was performed, showing extensive apical bone resorption on tooth #12 and dental resorption promoting communication of the root canal with the periodontium of tooth #11. The patient was referred to endodontic surgery. After surgery and postoperative periods of 15, 30, 60, 90, and 120days, standardized radiographs were taken, digitized, and subjected to digital subtraction of the images using Adobe Photoshop CS software. Four years later, a cone-beam computed tomography was performed, showing bone repair and absence of root resorption at tooth #11. In this clinical case, digital subtraction radiography was effective for early detection of new bone formation and evolution of repair. © 2011 John Wiley & Sons A/S.

Effects of a novel calcium aluminate cement on the early events of the progression of osteogenic cell cultures

The present study evaluated the progression of osteogenic cell cultures exposed to a novel calcium aluminate cement (CAC+) in comparison with the gold standard mineral trioxide aggregate (MTA). Cells were enzimatically isolated from newborn rat calvarial bone, plated on glass coverslips containing either CAC+ or a control MTA samples in the center, and grown under standard osteogenic conditions. Over the 10-day culture period, roundening of sample edges was clearly noticed only for MTA group. Although both cements supported osteogenic cell adhesion, spreading, and proliferation, CAC+-exposed cultures showed significantly higher values in terms of total cell number at days 3 and 7, and total protein content and alkaline phosphatase activity at day 10. The present in vitro results indicate that the exposure to CAC+ supports a higher differentiation of osteogenic cells compared with the ones exposed to MTA. Further experimental studies should consider CAC+ as a potential alternative to MTA when the repair of mineralized tissues is one of the desired outcomes in endodontic therapy.

Chronic draining mandibular fistula in a Californian Sea Lion

A male Californian Sea Lion with a chronic draining mandibular fistula had failed to respond to conventional treatments. This was effectively treated by endodontic therapy utilising calcium hydroxide to seal the apex and glass ionomer cement to obturate the root canal of the canine tooth.

The effect of timing temporary cements to treat induced pulp necrosis in the teeth of dogs

During endodontic therapy (pulpectomy, root canal debridement and root canal filling) microbiological management is a major concern. Bacteria present in dentine tubules, apical foramina and apical delta are causally related to failure of the procedure. Studies have shown that during single session endodontic treatment bacteria remain within dental structures. The aim of the present study was to evaluate endodontic treatment performed as two sessions, using temporary endodontic dressing materials for different periods in four groups of experimental dogs. A total of 80 roots of second and third upper premolar teeth and second, third and fourth lower premolar teeth were divided into four groups. The pulp chamber was opened with burrs and the pulp exposed for 60 days to induce pulpal inflammation and necrosis. Groups II, III and IV were treated with calcium hydroxide plus camphorated paramono-chlorophenol (PMCC) for 7, 15 and 30 days, respectively. In all groups, the root canals were filled with zinc oxide-eugenol and gutta-percha cones. Clinical and radiographical measurements were performed every 2 weeks. After 60 days a small block section containing the teeth, surrounding periapical tissues and the periodontium was removed for histological and microbiological study. Histological analysis revealed intense inflammatory response in all groups. Microbiological analysis showed microbial reduction inversely proportional to the period of time that the intracanal temporary medicament was left in place.

Inhibition of Sodium Hypochlorite Antimicrobial Activity in the Presence of Bovine Serum Albumin

Introduction: This study investigated the inhibition of the antimicrobial activity of sodium hypochlorite (NaOCl) by bovine serum albumin (BSA). The killing of Enterococcus faecalis, Candida albicans, Staphylococcus epidermidis, and Escherichia coil by NaOCl in concentrations from 2% to 0.03% was measured in the presence of BSA in concentrations between 6.7% and 0.1%. Methods: NaOCl, BSA, and microorganism suspensions were mixed, and, after 30 seconds, 6 minutes, and 30 minutes, samples were taken and NaOCl was inactivated by 5% sodium thiosulphate. The microbes were incubated in tryptic soy broth broth for up to 7 days for the detection of growth. Results: All microorganisms were killed within 30 seconds by 0.03% NaOCl when BSA was not present. High concentrations of BSA significantly reduced the antimicrobial activity of NaOCl against the four species. Conclusions: The inhibition of sodium hypochlorite by BSA was directly dependent on their quantitative relationships. The result partly explains the poorer performance in vivo of NaOCl as compared to in vitro experiments. (J Endod 2010;36:268-271)

Avaliacão da satisfação do usuário e da qualidade dos tratamentos endodônticos em Centros de Especialidades Odontológicas da Grande Natal-RN

Objective: To evaluate the degree of users satisfaction and technical quality of endodontic treatment in specialized dental clinics (CEO) of Grande Natal / RN between 2006 and 2008. Methodology: evaluated 282 endodontically treated teeth in CEOs through clinical and radiographic examinations. A questionnaire about the clinical condition of the tooth, evaluation of care and satisfaction with treatment was applied. Data on pre-and trans-operative were noted by the patient's clinical record. Endodontically treated teeth were examined by a specialist in endodontics, which compared with previous radiographs and current ones. The collected datas were presented descriptively by absolute numbers, percentages, averages. To determine the association between the independent and dependent variables was carried out through the bivariate association test Chi-square and Fisher exact test. Results: 79.8% presented with radiographic normal and 84.4% without pain symptoms. 8.2% of the teeth were fractured and 3.2% extracted. The persistence of the periapical lesion was associated with initial periapical status (p <0.05). 91.5% of patients are satisfied with the outcome of treatment. Such satisfaction is associated with absence of pain and an adequate esthetic tooth position (p <0.05). Conclusion: endodontic treatment in specialized dental clinics have an adequate technical quality, resulting in the success of endodontic therapy performed in these centers and that users are satisfied with the treatment